外周血单个核细胞甲胎蛋白mRNA表达与肝癌的研究.docVIP

外周血单个核细胞甲胎蛋白mRNA表达与肝癌的研究 肿瘤 1999年第6期第19卷 论著 作者：罗　伟　王　红　王　强　矢野右人 单位：罗伟　王红　王强　上海第二医科大学医学检验实验室(上海200023)；矢野右人　日本国立长崎中央病院临床研究部 关键词：甲胎蛋白；肝肿瘤/诊断；外周血单个核细胞；肿瘤转移/诊断 　　中图分类号：R735.7　　　文章标识码：A　　　文章编号：1000-7431(1999)06-0323-04 　　摘要：目的　应用RT-PCR技术研究原发性肝癌患者外周血单个核细胞(PBMC)甲胎蛋白(AFP)mRNA表达与肝癌转移的关系。方法　原发性肝癌48例。慢性肝病患者21例。正常对照20例。采外周血，肝素抗凝，分离单个核细胞及分离总RNA。逆转录酶合成cDNA。设计扩增AFP基因的引物，进行巢式PCR扩增。结果　正常人PBMC均无AFP mRNA表达。原发性肝癌患者的PBMC AFP mRNA阳性率(28/48，58.33%)明显高于慢性肝病组(3/21，14.28%，P＜0.01)。伴远处转移的原发性肝癌患者全部为阳性(8/8)。结论　RT-PCR检测原发性肝癌患者AFP mRNA有可能成为反映肝癌细胞血行播散的指标。 EXPRESSION OF AFP mRNA IN PERIPHERAL BLOOD CELLS OF PATIENTS WITH HEPATOCELLULAR CARCINOMA LUO Wei1，WANG Hong1,WANG Qiang1,et al. (1Laboratory of Medical Technology,Shanghai Second Medical University,Shanghai 200023,China；2Clinical Research Branch,National Nagasaki Central Hospital,Japan) 　　Abstract：Objective　A nested reverse transcription polymerase chain reaction(RT-PCR) was developed to study the relationship between expression of AFP mRNA in peripheral blood mononuclear cells and metastasis in patients with hepatocellular carcinoma(HCC).Methods Heparinized blood samples were obtained from 48 patients with HCC,21 patients with chronic liver diseases and 20 normal subjects.Mononuclear cells were separated from blood.Total RNA was extracted by the acid-guanidium-phenol-chloroform method.The first strand of cDNA was synthesized from an aliquot of the RNA using Moloney murine leukemia virus reverse transcriptase and random hexamers.The two pairs of primers chosen from the sequences of AFP were used for the two-step PCR.Results AFP mRNA in peripheral blood cells was not detected in all normal subjects, but it was detecte in 28 of 48 (58.33%) cases with HCC and in all 8 patients with extrahepatic metastasis.The positive rate in patients with HCC was significantly higher than the cases with chronic liver diseases(3/21,14.28%,P＜0.01).Conclusions The detection of AFP mRNA in peripheral blood cells may be helpful for clinial detection of metastasis o